Abstract
There is a strong need for generation and publication of reference values of immunotoxicity parameters within experimental and toxicological areas. This is particularly true where the type of distribution of reference values and thus the statistical method to be employed is often unknown and will become necessary if current official proposals to implement immune parameters into regulatory toxicity studies are adopted.
T and B lymphocytes were identified and quantitated by Fluorescence Activated Cell Sorter Analysis (FACS, using either one or two colours) after labelling of lymphocytes in suspension with fluorescent monoclonal antibodies (MoAbs). For single labelling of T and B lymphocytes, fluorescein isothiocyanate (FITC)-conjugated monoclonal antibody (MoAb) Ox-19 and Mark-1 were used respectively. In double labelling, T and B lymphocytes were identified with phycoerythrin (PE)-conjugated Ox-19 and FITC-conjugated Mark-1 MoAbs respectively. The physicochemical stability shelf-life of labelled T lymphocytes was at least 24 h at 2–8°C permitting overnight storage before FACS analysis, whereas enumeration of B lymphocytes should preferably be done directly after labelling. The sum of the percentages of T and B lymphocytes obtained in the combined T+B lymphocyte analysis with FITC-conjugated Ox-19 and Mark-1 were compared with the sum of values obtained in separate experiments after single labelling, yielding an inaccuracy of 0.3%. The precision of B and T cell analysis after double labelling was found to be 8.2% and 0.5% respectively.
Baseline data (reference/normal values) were obtained for rat T lymphocytes (63%–90%) and for B lymphocytes (7.5%–28%).
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Salemink, P., Doorstam, D., Maris, J. et al. Analytical aspects of FRCS (Fluorescence Activated Cell Sorter) identification of rat T and B peripheral lymphocytes in toxicity studies. Comparative Haematology International 2, 220–226 (1992). https://doi.org/10.1007/BF00216098
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DOI: https://doi.org/10.1007/BF00216098